Molecular analysis of rugosity in a <i>Vibrio cholerae</i> O1 El Tor phase variant

Yildiz, Fitnat H.; Liu, Xiaole S.; Heydorn, Arne; Schoolnik, Gary K.

doi:10.1111/j.1365-2958.2004.04154.x

Cited by 237 publications

(373 citation statements)

References 51 publications

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“…Consistent with the biofilm data, the Dcrp mutant expressed elevated levels of the exopolysaccharide biosynthesis gene vpsL. VpsL has been shown to be repressed by HapR and contains a putative HapR binding site (Yildiz et al, 2004). However, Dcrp and DhapRcrp mutants made less vpsL mRNA than DhapR, suggesting that CRP could affect other factors required for maximal exopolysaccharide expression.…”

Section: Discussionsupporting

confidence: 72%

“…Interestingly, flaACD and flgL have also been found to be downregulated in a DhapR mutant (Yildiz et al, 2004). To determine the contribution of cAMP-CRP and HapR to the regulation of flagellar genes and motility we confirmed the downregulation of flaA and flaC by qRT-PCR and compared expression of flaA and flaC in wild-type, DhapR, Dcrp and DhapRcrp backgrounds.…”

Section: Effect Of Crp On Motilitymentioning

confidence: 67%

“…ToxT acts positively at the ctxA and tcpA promoters to activate production of cholera toxin (CT) and the toxin coregulated pilus (TCP) (DiRita et al 1991). At high cell density, HapR represses the transcription of aphA to diminish CT and TCP production (Kovacikova & Skorupski, 2002), represses the exopolysaccharide genes involved in biofilm formation (Yildiz et al, 2004) and activates hapA (Jobling & Holmes, 1997;Zhu et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

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The cyclic AMP receptor protein modulates quorum sensing, motility and multiple genes that affect intestinal colonization in Vibrio cholerae

et al. 2007

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Vibrio cholerae is the causative agent of cholera, which continues to be a major public health concern in Asia, Africa and Latin America. The bacterium can persist outside the human host and alternates between planktonic and biofilm community lifestyles. Transition between the different lifestyles is mediated by multiple signal transduction pathways including quorum sensing. Expression of the Zn-metalloprotease haemagglutinin (HA)/protease is subject to a dual regulation which involves the quorum-sensing regulator HapR and the cAMP receptor protein. In a previous study, we observed that a mutant defective in the cAMP-receptor protein (CRP) expressed lower levels of HapR. To further investigate the role of CRP in modulating HapR and other signal transduction pathways, we performed global gene expression profiling of a Dcrp mutant of El Tor biotype V. cholerae. Here we show that CRP is required for the biosynthesis of cholera autoinducer 1 (CAI-1) and affects the expression of multiple HapR-regulated genes. As expected, the Dcrp mutant produced more cholera toxin and enhanced biofilm. Expression of flagellar genes, reported to be affected in DhapR mutants, was diminished in the Dcrp mutant. However, an epistasis analysis indicated that cAMP-CRP affects motility by a mechanism independent of HapR. Inactivation of crp inhibited the expression of multiple genes reported to be strongly induced in vivo and to affect the ability of V. cholerae to colonize the small intestine and cause disease. These genes included ompU, ompT and ompW encoding outer-membrane proteins, the alternative sigma factor s E required for intestinal colonization, and genes involved in anaerobic energy metabolism. Our results indicate that CRP plays a crucial role in the V. cholerae life cycle by affecting quorum sensing and multiple genes required for survival of V. cholerae in the human host and the environment.

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Section: Discussionsupporting

confidence: 72%

Section: Effect Of Crp On Motilitymentioning

confidence: 67%

Section: Introductionmentioning

confidence: 99%

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The cyclic AMP receptor protein modulates quorum sensing, motility and multiple genes that affect intestinal colonization in Vibrio cholerae

et al. 2007

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“…A DhapR DflaA double deletion mutant (Nadell and Bassler, 2011), which secretes copious extracellular matrix, was used to produce resident biofilms. We refer to this strain as 'Rugose' (Yildiz et al, 2004), as it forms wrinkled colonies on agar. Invading strain genotypes were:…”

Section: Strainsmentioning

confidence: 99%

Extracellular matrix structure governs invasion resistance in bacterial biofilms

et al. 2015

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Many bacteria are highly adapted for life in communities, or biofilms. A defining feature of biofilms is the production of extracellular matrix that binds cells together. The biofilm matrix provides numerous fitness benefits, including protection from environmental stresses and enhanced nutrient availability. Here we investigate defense against biofilm invasion using the model bacterium Vibrio cholerae. We demonstrate that immotile cells, including those identical to the biofilm resident strain, are completely excluded from entry into resident biofilms. Motile cells can colonize and grow on the biofilm exterior, but are readily removed by shear forces. Protection from invasion into the biofilm interior is mediated by the secreted protein RbmA, which binds mother-daughter cell pairs to each other and to polysaccharide components of the matrix. RbmA, and the invasion protection it confers, strongly localize to the cell lineages that produce it.

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“…In Pseudomonas spp., the GacS/GacA system regulates phase variation to a Tr morphology that enhances host colonization (Han et al, 1997;Sánchez-Contreras et al, 2002). Phase variation to the rugose phenotype in V. cholerae enhances biofilm formation and is mediated through HapR (Yildiz et al, 2004), which is regulated by GacA input to the quorumsensing system (Lenz et al, 2005). Furthermore, GacAmediated switching in V. fischeri facilitates a symbiotic lifestyle (Whistler et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

Role of GacA in virulence of Vibrio vulnificus

et al. 2010

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The GacS/GacA two-component signal transduction system regulates virulence, biofilm formation and symbiosis in Vibrio species. The present study investigated this regulatory pathway in Vibrio vulnificus, a human pathogen that causes life-threatening disease associated with the consumption of raw oysters and wound infections. Small non-coding RNAs (csrB1, csrB2, csrB3 and csrC) commonly regulated by the GacS/GacA pathway were decreased (P,0.0003) in a V. vulnificus CMCP6 DgacA : : aph mutant compared with the wild-type parent, and expression was restored by complementation of the gacA deletion mutation in trans. Of the 20 genes examined by RT-PCR, significant reductions in the transcript levels of the mutant in comparison with the wildtype strain were observed only for genes related to motility (flaA), stationary phase (rpoS) and protease (vvpE) (P50.04, 0.01 and 0.002, respectively). Swimming motility, flagellation and opaque colony morphology indicative of capsular polysaccharide (CPS) were unchanged in the mutant, while cytotoxicity, protease activity, CPS phase variation and the ability to acquire iron were decreased compared with the wild-type (P,0.01). The role of gacA in virulence of V. vulnificus was also demonstrated by significant impairment in the ability of the mutant strain to cause either skin (P,0.0005) or systemic infections (P,0.02) in subcutaneously inoculated, noniron-treated mice. However, the virulence of the mutant was equivalent to that of the wild-type in iron-treated mice, demonstrating that the GacA pathway in V. vulnificus regulates the virulence of this organism in an iron-dependent manner. INTRODUCTIONVibrio vulnificus is a moderately halophilic, Gram-negative bacterium that inhabits coastal waters and colonizes fish and filter-feeding shellfish (DePaola et al., 1994;Motes et al., 1998;Tamplin & Capers, 1992; Wright et al., 1996). Individuals with underlying conditions such as liver disease, diabetes mellitus, cancer, haemochromatosis (iron-overload) or immune system dysfunction can incur life-threatening systemic disease from consumption of raw oysters or from wound infections (Blake et al., 1979;Jones & Oliver, 2009). The virulence of this bacterium in animal models has been attributed to multiple factors (Gulig et al., 2005;Jones & Oliver, 2009), including capsular polysaccharide (CPS) expression (Amako et al., 1984;Kreger et al., 1984;Simpson et al., 1987;Yoshida et al., 1985), iron acquisition (Litwin et al., 1996;Simpson & Oliver, 1983;Wright et al., 1986), pili (Gander & LaRocco, 1989;Paranjpye & Strom, 2005;Paranjpye et al., 2007), flagella (Kim & Rhee, 2003;Lee et al., 2004), quorum sensing (McDougald et al., 2001) and cytolytic haemolysin encoded by the rtxA1 gene (Lee et al., 2007; Kim et al., 2008;Liu et al., 2007). Other cytotoxin/haemolysin genes, including rtxA2 and rtxA3 (J. L. Joseph and P. A. Gulig, unpublished results) and vvhA (Wright & Morris, 1991), as well as a metalloprotease encoded by vvpE (Jeong et al., 2000;Shao & Hor, 2000), show no apparent role in the vi...

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Molecular analysis of rugosity in a Vibrio cholerae O1 El Tor phase variant

Cited by 237 publications

References 51 publications

The cyclic AMP receptor protein modulates quorum sensing, motility and multiple genes that affect intestinal colonization in Vibrio cholerae

The cyclic AMP receptor protein modulates quorum sensing, motility and multiple genes that affect intestinal colonization in Vibrio cholerae

Extracellular matrix structure governs invasion resistance in bacterial biofilms

Role of GacA in virulence of Vibrio vulnificus

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